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10.1128/JVI.01265-20

http://scihub22266oqcxt.onion/10.1128/JVI.01265-20
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32843534!7592234!32843534
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suck abstract from ncbi


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pmid32843534      J+Virol 2020 ; 94 (22): ä
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  • Development of a Fluorescence-Based, High-Throughput SARS-CoV-2 3CL(pro) Reporter Assay #MMPMID32843534
  • Froggatt HM; Heaton BE; Heaton NS
  • J Virol 2020[Oct]; 94 (22): ä PMID32843534show ga
  • In late 2019, a human coronavirus, now known as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), emerged, likely from a zoonotic reservoir. This virus causes COVID-19, has infected millions of people, and has led to hundreds of thousands of deaths across the globe. While the best interventions to control and ultimately stop the pandemic are prophylactic vaccines, antiviral therapeutics are important to limit morbidity and mortality in those already infected. At this time, only one FDA-approved anti-SARS-CoV-2 antiviral drug, remdesivir, is available, and unfortunately, its efficacy appears to be limited. Thus, the identification of new and efficacious antivirals is of the highest importance. In order to facilitate rapid drug discovery, flexible, sensitive, and high-throughput screening methods are required. With respect to drug targets, most attention is focused on either the viral RNA-dependent RNA polymerase or the main viral protease, 3CL(pro) 3CL(pro) is an attractive target for antiviral therapeutics, as it is essential for processing newly translated viral proteins and the viral life cycle cannot be completed without protease activity. In this work, we report a new assay to identify inhibitors of 3CL(pro) Our reporter is based on a green fluorescent protein (GFP)-derived protein that fluoresces only after cleavage by 3CL(pro) This experimentally optimized reporter assay allows for antiviral drug screening in human cell culture at biosafety level 2 (BSL2) with high-throughput compatible protocols. Using this screening approach in combination with existing drug libraries may lead to the rapid identification of novel antivirals to suppress SARS-CoV-2 replication and spread.IMPORTANCE The COVID-19 pandemic has already led to more than 700,000 deaths and innumerable changes to daily life worldwide. Along with development of a vaccine, identification of effective antivirals to treat infected patients is of the highest importance. However, rapid drug discovery requires efficient methods to identify novel compounds that can inhibit the virus. In this work, we present a method for identifying inhibitors of the SARS-CoV-2 main protease, 3CL(pro) This reporter-based assay allows for antiviral drug screening in human cell culture at biosafety level 2 (BSL2) with high-throughput compatible sample processing and analysis. This assay may help identify novel antivirals to control the COVID-19 pandemic.
  • |*Drug Discovery[MESH]
  • |Animals[MESH]
  • |Antiviral Agents/*pharmacology[MESH]
  • |Betacoronavirus/*chemistry[MESH]
  • |COVID-19[MESH]
  • |Chlorocebus aethiops[MESH]
  • |Coronavirus 3C Proteases[MESH]
  • |Coronavirus Infections/drug therapy/*virology[MESH]
  • |Cysteine Endopeptidases[MESH]
  • |High-Throughput Screening Assays/*methods[MESH]
  • |Humans[MESH]
  • |Microscopy, Fluorescence/methods[MESH]
  • |Pandemics[MESH]
  • |Pneumonia, Viral/drug therapy/*virology[MESH]
  • |Protease Inhibitors/*pharmacology[MESH]
  • |SARS-CoV-2[MESH]
  • |Vero Cells[MESH]


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